Nucleoside peptide antibiotics of AA-896

ABSTRACT

This invention relates to antibiotic compounds AA896 of the formulawherein:R1 is H, aryl, alkyl (C1-C20), -CH2-aryl, -C(O)alkyl(C1-C20), -C(O)NHalkyl(C1-C20), or -C(O)NHaryl;R2 is H, alkyl (C1-C20), -CH2aryl, alkyl (C1-C20) or -C(O)alkyl(C1-C20);R3 is -OH;R2 and R3 may optionally be taken together to form a moietyR4 is alkyl (C1-C20), or aryl;or a pharmaceutically acceptable salt thereof.

CROSS REFERENCE TO RELATED APPLICATIONS

“This application claims priority from copending provisionalapplications Serial No. 60/286,401 filed on Apr. 25, 2001, Serial No.60/290,139, filed on May 10, 2001, Serial No. 60/286,297 filed Apr. 25,2001, Serial No. 60/290,140 filed May 10, 2001, Serial No. 60/286,402filed Apr. 25, 2001 and Serial No. 60/290,156 filed May 10, 2001 theentire disclosures of which are hereby incorporated by reference.”

FIELD OF THE INVENTION

The present invention relates to novel compounds of AA-896 which exhibitantibacterial activity.

BACKGROUND OF THE INVENTION

Natural products, Liposidomycins A, B and C, have been isolated andreported to have antibacterial activity(Isono, K.; Uramoto, M.;Kusakabe, H.; Kimura, K.; Izaki, K.; Nelson, C. C.; McCloskey, J. A.,J.Antibiotics, 1985, 38, 1617-1621. Ubukata, M.; Isono, K.; Kimura, K.;Nelson, C. C.; McCloskey, J. A. J.Am.Chem.Soc., 1988,110, 4416-4417.Kimura, K.; Miyata, N.; Kawanishi, G.; Kamino, Y.; Izaki, K.; Isono, K.Agric.Biol.Chem., 1989, 53,1811-1815.). Isolated Liposidomycins A-(I),A-(II), A-(III) and A-(IV) are also reported to have antibacterialactivity (Kimura, K.; Ikeda, Y.; Kagami, S.; Yoshihara, M., J.Antibiotics, 1998, 51, 1099-1104. and other references herein). Thedetailed structural analysis of Liposidomycins A, B and C using theirchemical degradation products has been reported (Ubukata, M.; Kimura,K.; Isono, K.; Nelson, C. C.; Gregson, J. M.; McClosky, J. A.,J.Org.Chem., 1992, 57, 6392-6403). Liposidomycin class compounds arefurther reported (JPO05078385) and are derivatives of2-methylamino-3-(5-aminomethyl-4-hydroxy-3-hydroxy-tetrahydro-fura-2-yloxy)-3-[3,4-dihyoxy-5-(2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl)tetrahydro-2-furanyl]propanoic acid or the degradedproducts of2-methylamino-3-(5-aminomethyl-4-hydroxy-3-hydroxy-tetrahydro-fura-2-yloxy)-3-[3,4-dihyoxy-5-(2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl)tetrahydro-2-furanyl]-propanoic acid.

This invention is concerned with a new series of nucleoside peptideantibiotics of AA-896.

SUMMARY OF THE INVENTION

This invention is concerned with novel nucleoside peptide antibiotics ofAA-896 which have antibacterial activity; with methods of treatinginfectious disease in mammals employing these novel nucleoside peptideantibiotics; with pharmaceutical compositions containing these novelnucleoside peptide antibiotics and processes for the production of novelnucleoside peptide antibiotics of the invention. Compounds according tothe invention comprise compounds of the formula

wherein:

R₁ is H, aryl, alkyl (C₁-C₂₀), —CH₂-aryl, —C(O)alkyl(C₁-C₂₀),C(O)NHalkyl(C₁-C₂₀), or —C(O)NHaryl;

R₂ is H, alkyl (C₁-C₂₀), —CH₂aryl, or —C(O)alkyl(C₁-C₂₀);

R₃ is —OH;

R₂ and R₃ may optionally be taken together to form a moiety

R₄ is alkyl (C₁-C₂₀), or aryl;

provided R₁ and R₂ are not H when R₃ is —OH

or a pharmaceutically acceptable salt thereof.

Among the preferred groups of compounds of this invention includingpharmaceutically acceptable salts thereof are those in the subgroupsbelow, wherein other variables are as defined above:

a) R₂ and R₃ are taken together to form a moiety

b) R₂ is H, alkyl (C₁-C₁₂), or —CH₂aryl; and

c) R₁ is H, —C(O)alkyl(C₁-C₁₆), or —C(O)aryl when

R₂ and R₃ are taken together to form a moiety

d) R₄ is alkyl(C₁-C₁₆), or aryl.

Specifically preferred compounds of the invention are the followingcompounds or a pharmaceutically acceptable salt thereof:

14-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-3-(4-fluorophenyl)-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid,

14-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-2,4-dioxo-3-pentyl-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid,

14-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{((2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-3-hexyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid,

14-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-3-(4-methoxyphenyl)-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid,

14-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-3-dodecyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid,

16-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-15-benzyl-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid,

16-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-15-dodecyl-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid,

16-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-15-[12-(4-morpholinyl)dodecyl]-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid,

16-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-15-pentyl-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid,

4-[13-((2R)-2-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}-1-carboxy-2-{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}ethyl)-26-carboxy-19-(1-hydroxy-2-methylpropyl)-22-(2-iminohexahydro-4-pyrimidinyl)-27-methyl-18,21,24-trioxo-13,17,20,23,25-pentaazaoctacos-1-yl]-4-methylmorpholin-4-ium,

14-[5-((R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(3R,4S,5R)-4-hydroxy-3-methoxy-5-({[(octylamino)carbonyl]amino}methyl)tetrahydro-2-furanyl]oxy}methyl)-3-octyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid,

14-[5-((R)-{(2S,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(5R)-4-hydroxy-3-methoxy-5-({[(4-fluoroanilino)carbonyl]amino}methyl)tetrahydro-2-furanyl]oxy}methyl)-3-(4-fluorophenyl)-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid,

14-[5-((R)-{(2S,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(5R)-4-hydroxy-3-methoxy-5-({[(4-methoxyanilino)carbonyl]amino}methyl)tetrahydro-2-furanyl]oxy}methyl)-3-(4-methoxyphenyl)-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid,

14-[5-((R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(3R,4S,5R)-5-({[(hexylamino)carbonyl]amino}methyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}methyl)-3-hexyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid,

14-[5-((R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(3R,4S,5R)-5-({[(dodecylamino)carbonyl]amino}methyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}methyl)-3-dodecyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid,

16-((R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(3R,4S,5R)-5-({[(hexadecylamino)carbonyl]amino}methyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid,

16-[(R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}({(3R,4S,5R)-4-hydroxy-3-methoxy-5-[(pentylamino)methyl]tetrahydro-2-furanyl}oxy)methyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-15-pentyl-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid,

16-[(R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}({(3R,4S,5R)-4-hydroxy-3-methoxy-5-[(pentylamino)methyl]tetrahydro-2-furanyl}oxy)methyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid,

16-((R)-[((3R,4S,5R)-5-{[([1,1′-Biphenyl]-4-ylmethyl)amino]methyl}-4-hydroxy-3-methoxytetrahydro-2-furanyl)oxy]{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid,

15-([1,1′-Biphenyl]-4-ylmethyl)-16-((R)-[((3R,4S,5R)-5-{[([1,1′-biphenyl]-4-ylmethyl)amino]methyl}-4-hydroxy-3-methoxytetrahydro-2-furanyl)oxy]{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid,

16-((R)-({(3R,4S,5R)-5-[(Benzylamino)methyl]-4-hydroxy-3-methoxytetrahydro-2-furanyl}oxy){(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid,

15-Benzyl-16-((R)-({(3R,4S,5R)-5-[(benzylamino)methyl]-4-hydroxy-3-methoxytetrahydro-2-furanyl}oxy){(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid, and

16-((R)-({(3R,4S,5R)-5-[(Acetamido)methyl]-4-hydroxy-3-methoxytetrahydro-2-furanyl}oxy){(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid.

For the compounds of the invention defined above and referred to herein,unless otherwise noted, the following terms are defined:

Halogen, as used herein means fluoro, chloro, bromo and/or iodo.

Alkyl as used herein means a branched or straight chain radical havingfrom 1 to 20 (preferably 1 to 16) or preferably (1 to 12)carbon atomsoptionally substituted with morpholino where the morpholino nitrogenatom may optionally be alkylated with alkyl of 1 to 6 carbon atomsforming a quaternary salt. Exemplary alkyl groups include but are notlimited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl,pentyl and hexyl, also optionally substituted, as well asperfluoroalkyl.

Aryl as used herein means a homocyclic or polycyclic aromatic radical,having 6 to 20 carbon atoms independently substituted with one to threesubstituents selected from the group of alkyl, halogen, cyano, nitro,hydroxy, , amino, alkylamino, dialkylamino, or alkoxy. Examples include,but are not limited to, phenyl, biphenyl, naphthyl, fluorenyl, andanthracenyl, optionally substituted with one to three substituents.

The preparation of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid by fermentation is described in copending application Ser. No.60/286,402 filed Apr. 25, 2001 incorporated herein by reference andcopending application Ser. No. 60/290,156 filed May 10, 2001incorporated herein by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Proton NMR spectrum of Example 1 in D₂O at 300 MHz

FIG. 2. Proton NMR spectrum of Example 5 in D₂O at 300 MHz

FIG. 3. Proton NMR spectrum of Example 6 in D₂O at 300 MHz

FIG. 4. Proton NMR spectrum of Example 7 in D₂O at 300 MHz

FIG. 5. Proton NMR spectrum of Example 8 in D₂O at 300 MHz

FIG. 6. Proton NMR spectrum of Example 9 in D₂O at 300 MHz

FIG. 7. Proton NMR spectrum of Example 10 in D₂O at 300 MHz

FIG. 8. Proton NMR spectrum of Example 11 in D₂O at 300 MHz

FIG. 9. Proton NMR spectrum of Example 12 in D₂O at 300 MHz

FIG. 10. Proton NMR spectrum of Example 14 in D₂O at 300 MHz

FIG. 11. Proton NMR spectrum of Example 16 in D₂O at 300 MHz

FIG. 12. Proton NMR spectrum of Example 18 in D₂O at 300 MHz

FIG. 13. Proton NMR spectrum of Example 19A in D₂O at 300 MHz

FIG. 14. Proton NMR spectrum of Example 19B in D₂O at 300 MHz

FIG. 15. Proton NMR spectrum of Example 20A in D₂O at 300 MHz

FIG. 16 Proton NMR spectrum of Example 20B in D₂O at 300 MHz

FIG. 17 Proton NMR spectrum of Example 24 in D₂O at 300 MHz

DETAILED DESCRIPTION OF INVENTION

The compounds of the invention are prepared according to the followingreaction schemes.

As shown in Scheme 1, protecting the primary amine of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid 1 with 2,4-pentanedione in pyridine and methanol is followed byreaction with an isocyanate R₄NCO 2 in a solvent such asN,N-dimethylformamide with later removal of the amine protecting groupusing 0.5% trifluoroacetic acid in water and dioxane to give carboxylicacid 3.

In accordance with Scheme 2,16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid 1 is reacted with 2,4-pentanedione, to protect the primary amine,followed by reaction with arylCH₂Br 4 in a solvent such asN,N-dimethylformamide followed by removal of the protecting group with0.5% trifluoroacetic acid in water and dioxane affords dicarboxylic acid5.

As shown in Scheme 3, reaction of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid 1 having the primary amine protected with 2,4-pentanedione followedby reductive alkylation by reaction with R₄CHO 6 in a solvent such asN,N-dimethylformamide followed by treatment with either sodiumcyanoborohydride or sodium triacetoxyborohydride and then removal of theprimary amine protecting group with 0.5% trifluoroacetic acid in waterand dioxane affords alkylated derivatives 7.

As shown in Scheme 4, reaction of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid 1 with R₄NCO 2 in a solvent such as N,N-dimethylformamide followedby treatment with water affords dialkylated derivatives 8.

In accordance with Scheme 5, reaction of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid 1, by reductive alkylation with R₄CHO 6 in a solvent such asN,N-dimethylformamide followed by treatment with either sodiumcyanoborohydride or sodium triacetoxyborohydride affords dialkylatedderivatives 9 or monoalkylated derivatives 9a.

As shown in Scheme 6, reaction of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid 1, with acetic anhydride (Ac₂O) at room temperature affordsderivative 10.

It is understood that compounds of this invention encompasses allcrystalline forms. Further, compounds of the invention may be obtainedas pharmaceutically acceptable salts which are those derived from suchorganic and inorganic acids as:

acetic, trifluoroacetic, lactic, citric, tartaric, formate, succinic,maleic, malonic, gluconic, hydrochloric, hydrobromic, phosphoric,nitric, sulfuric, methanesulfonic, and similarly known acceptable acids.Additionally, compounds of the invention may form calcium, potassium,magnesium, or sodium salts. The pharmaceutically acceptable salts ofcompounds of the invention are prepared using conventional procedures.Compounds of the invention have centers of asymmetry. The compounds ofthe invention may, therefore, exist in at least two and often morestereoisomeric forms. The present invention encompasses allstereoisomers whether free from other stereoisomers or admixed withother stereoisomers in any proportion and thus includes, for instance,racemic mixtures of enantiomers as well as the diastereomeric mixturesof isomers. The absolute configuration of any substantially purecompound may be determined by any suitable method including conventionalX-ray crystallography.

The present invention accordingly provides a pharmaceutical compositionwhich comprises a compound of the invention or a mixture thereof incombination or association with a pharmaceutically acceptable carrier.In particular, the present invention provides a pharmaceuticalcomposition which comprises an effective amount of compounds of theinvention or a mixture thereof and a pharmaceutically acceptablecarrier.

The present invention also provides methods which may be used intreating bacterial infections in warm blooded animals which compriseproviding to said animals an antibacterially effective amount of acompound of the invention or a mixture thereof.

Biological Activity

In Vitro Evaluation of AA-896 Derivatives as Antibacterial Agents

The in vitro antibacterial activity of Example 1 through Example 20 isdetermined against a spectrum of Gram-positive and Gram-negativebacteria by a standard broth dilution method. Serial dilution of thecompounds are made in Mueller-Hinton broth and inoculated with abacterial suspension. The lowest concentration of compound thatinhibited the growth of a bacterial strain after 18 hours of incubationat 35° C. is reported as the minimal inhibitory concentration (MIC) forthat strain. The results are given in Table 1.

TABLE 1 Antimicrobial Activity (MIC, μg/mL) of AA-896 DerivativesExample Example Example Example Example Example 1 2 3 4 5 6 E. coli GC4559 >32 >32 >32 >32 >32 >32 E. coli GC 4560  64  32  32 >32  4  2 E.coli GC 3226 >32 >32 >32 >32 >32 >32 S. marcescensGC >32 >32 >32 >32 >32 >32 4077 P. rettgeri GC4530 >32 >32 >32 >32 >32 >32 M. morganii GC 4531 >32 >32 >32 >32 >32 >32K. pneumoniae GC >32 >32 >32 >32 >32 >32 4534 E. cloacae GC3783 >32 >32 >32 >32 >32 >32 P. aeruginosa GC >32 >32 >32 >32 >32 >324532 S. aureus GC 4536 >32 >32 >32 >32 >32 >32 S. aureus GC1131 >32 >32 >32 >32 >32 >32 CNS GC 4537 >32 >32 >32 >32 >32 >32 CNS GC4538  32 >32 >32 >32 >32 >32 CNS GC 4547 >32 >32 >32 >32 >32 >32 E.faecalis GC 842 >32 >32 >32 >32 >32 >32 E. faecalis GC2242 >32 >32 >32 >32 >32 >32 E. coli GC 2203 >32 >32 >32 >32 >32 >32 P.aeruginosa GC >32 >32 >32 >32 >32 >32 2214 S. aureus GC2216 >32 >32 >32 >32 >32 >32 E. faecalis GC 4555 >32 >32 >32 >32 >32 >32Example Example Example Example Example Example Example 7 8 9 10 11 1213 E. coil GC 4559 >32 >32 >32 >32 >32 >32 >32 E. coli GC 4560  16 >32 32  16 >32 >32 >32 E. coli GC 3226 >32 >32 >32 >32 >32 >32 >32 S.marcescens >32 >32 >32 >32 >32 >32 >32 GC 4077 P. rettgeriGC >32 >32 >32 >32 >32 >32 >32 4530 M. morganiiGC >32 >32 >32 >32 >32 >32 >32 4531 K.pneumoniae >32 >32 >32 >32 >32 >32 >32 GC 4534 E. cloacaeGC >32 >32 >32 >32 >32 >32 >32 3783 P.aeruginosa >32 >32 >32 >32 >32 >32 >32 GC 4532 S. aureusGC >32 >32 >32 >32 >32 >32 >32 4536 S. aureusGC >32 >32 >32 >32 >32 >32 >32 1131 CNS GC4537 >32 >32 >32 >32 >32 >32 >32 CNS GC 4538 >32 >32 >32 >32 >32 >32 >32CNS GC 4547 >32 >32 >32 >32 >32 >32 >32 E. faecalisGC >32 >32 >32 >32 >32 >32 >32 842 E. faecalisGC >32 >32 >32 >32 >32 >32 >32 2242 E. coli GC2203 >32 >32 >32 >32 >32 >32 >32 P.aeruginosa >32 >32 >32 >32 >32 >32 >32 GC 2214 S. aureusGC >32 >32 >32 >32 >32 >32 >32 2216 E. faecalisGC >32 >32 >32 >32 >32 >32 >32 4555 Example Example Example ExampleExample 14 15 16 17 18 E. coli GC 4559 >32 >32 >32 >32 >32 E. coli GC4560  32 >32 >32  32 >32 E. coli GC 3226 >32 >32 >32 >32 >32 S.marcescens GC > 32 > 32 > 32 > 32 > 32 4077 P. rettgeri GC4530 >32 >32 >32 >32 >32 M. morganii GC 4531 > 32 > 32 > 32 > 32 > 32 K.pneumoniae GC >32 >32 >32 >32 >32 4534 E. cloacae GC3783 >32 >32 >32 >32 >32 P. aeruginosa GC 4532 > 32 > 32 >32 > 32 > 32S. aureus GC 4536  32 >32 >32 >32 >32 S. aureus GC 1131 32 >32 >32 >32 >32 CNS GC 4537  32 >32 >32 >32 >32 CNS GC 4538 32 >32 >32 >32 >32 CNS GC 4547  16 >32 >32  32 >32 E. faecalis GC842 >32 >32 >32 >32 >32 E. faecalis GC 2242  32 >32 >32  32 >32 E. coliGC 2203 >32 >32 >32 >32 >32 P. aeruginosa GC 2214 >32 >32 >32 >32 >32 S.aureus GC 2216  32 >32 >32 >32 >32 E. faecalis GC4555 >32 >32 >32 >32 >32 Example Example Example Example 19A 19B 20A 20BE. coli GC 4559 >32 >32 >32 >32 E. coli GC 4560  16 >32 >32  32 E. coliGC 3226 >32 >32 >32 >32 S. marcescens GC 4077 >32 > 32 > 32 > 32 P.rettgeri GC 4530 >32 >32 > 32 >32 M. morganii GC 4531 >32 >32 >32 >32 K.pneumoniae GC 4534 > 32 > 32 >32 >32 E. cloacae GC 3783 >32 > 32 >32 >32P. aeruginosa GC 4532 >32 >32 >32 >32 S. aureus GC 4536 >32 >32 >32 >32S. aureus GC 1131 >32 >32 >32 >32 CNS GC 4537 >32 >32 >32 >32 CNS GC4538 >32 >32 >32 >32 CNS GC 4547 >32 >32 >32 >32 E. faecalis GC842 >32 >32 >32 >32 E. faecalis GC 2242 >32 >32 >32 >32 E. coli GC2203 >32 >32 >32 >32 P. aeruginosa GC 2214 >32 >32 >32 >32 S. aureus GC2216 >32 >32 >32 >32 E. faecalis GC 4555 >32 > 32 > 32 > 32

Determination of Lipid II Formation by a TLC Methodology

The MurG biochemical assay utilizes S. epidermides membranes to catalyzethe late steps in cell wall biosynthesis including MraY, thephospho-MurNAc pentapeptide translocase and MurG, theUDP-N-Acetylglucosaminyl transferase. The following procedure is adaptedfrom the method described by Mengin-Lecreaulx, et al (J. Bacteriol173(15) 4625-4636, 1991). In this procedure, the formation of Lipid IIis assessed using radiolabeled UDP-N-Acetylglucosamine.

S. epidermides membranes, compound, UDP-MurNAc pentapeptide, and[¹⁴C]-UDP-N-Acetylglucosamine were incubated at room temperature for 30min. The reaction was terminated by boiling in a water bath for 1minute. 2 μl samples of each reaction are analyzed by TLC. The samplesare spotted onto K6 silica plates and chromatographed in IsobutyricAcid:1 M NH₄OH (5:3). The plates are exposed to film and the inhibitionof Lipid II formation can be monitored by comparing the sample area tothe control area.

IC₅₀ 100 μg/ml 50 μg/ml 25 μg/ml 6.25 μg/ml Example 1 + + + − Example2 + − − − Example 5 + + + + Example 6 + + + + Example 7 + + − − Example8 + − − − Example 9 + − − − Example 11 − − − − Example 12 − − − −Example 13 − − − − Example 14 − − − − Example 15 − − − − Example 16 + −− − Example 17 − − − − Example 19A + + + − Example 19B − − − −

Compounds of the invention derive their utility from their antibacterialactivity. For example, these compounds may be used in the suppression ofbacterial infections, as a topical antibacterial agent and as a generaldisinfectant for laboratories.

In therapeutic use, the compounds of this invention may be administeredin the form of conventional pharmaceutical compositions appropriate forthe intended use. Such compositions may be formulated so as to besuitable for oral, parenteral or topical administration. The compoundsof the invention may be combined in admixture with a nontoxicpharmaceutical carrier, which carrier may take a variety of forms,depending on the form of preparation desired for administration, ie.oral, parenteral, or topical.

When the compounds of the invention are employed for the above utility,they can be combined with one or more pharmaceutically acceptablecarriers, for example, solvents, diluents and the like, and may beadministered orally in such forms as tablets, capsules, dispersiblepowders, granules, or suspensions containing, for example, from about0.05 to 5% of suspending agent, syrups containing, for example fromabout 10 to 50% of sugar, and elixirs containing, for example from about20 to 50% ethanol, and the like, or parenterally in the form of sterileinjectable solutions or suspensions containing from about 0.05 to 5%suspending agent in an isotonic medium. Such pharmaceutical preparationsmay contain, for example, from about 0.05 up to about 90% of thecompounds of the invention in combination with the carrier, more usuallybetween about 5% and 60% by weight.

An antibacterially effective amount of compounds of the invention fromabout 0.5 mg/kg of body weight to about 200.0 mg/kg of body weightshould be administered one to five times per day via any topical routesof administration including but not limited to oral, parenteral(including subcutaneous, intravenous, intramuscular, intrasternalinjection or infusion techniques), by inhalation spray, or rectally, indosage unit formulations containing conventional non-toxicpharmaceutically acceptable carriers, adjuvants and vehicles. It will beunderstood, however, that the specific dose level and frequency ofdosage for any particular patient may be varied and will depend upon avariety of factors including the activity of the specific compoundemployed, the metabolic stability and length of action of that compound,the age, body weight, general health, sec, diet, mode and time ofadministration, rate of excretion, drug combination, the severity of theparticular condition, and the host undergoing therapy.

Additionally, the antibacterially effective amount of the compounds ofthe invention may be administered at a dosage and frequency withoutinducing side effects commonly experienced with conventional antibiotictherapy which could include hypersensitivity, neuromuscular blockade,vertigo, photosensitivity, discoloration of teeth, hematologic changes,gastrointestinal disturbances, ototoxicity, and renal, hepatic, orcardiac impairment. Further the frequency and duration of dosage may bemonitored to substantially limit harmful effects to normal tissuescaused by administration at or above the antibacterially effectiveamount of the substantially pure compounds of the invention.

These active compounds may be administered orally as well as byintravenous, intramuscular, or subcutaneous routes. Solid carriersinclude starch, lactose, dicalcium phosphate, microcrystallinecellulose, sucrose and kaolin, while liquid carriers include sterilewater, polyethylene glycols, non-ionic surfactants and edible oils suchas corn, peanut and sesame oils, as are appropriate to the nature of theactive ingredient and the particular form of administration desired.Adjuvants customarily employed in the preparation of pharmaceuticalcompositions may be advantageously included, such as flavoring agents,coloring agents, preserving agents, and antioxidants, for example,vitamin E, ascorbic acid, BHT and BHA. The preferred pharmaceuticalcompositions from the stand-point of ease of preparation andadministration are solid compositions, particularly tablets andhard-filled or liquid-filled capsules. Oral administration of thecompound is preferred. The compounds of the invention may also beadministered parenterally or intraperitoneally. Solutions or suspensionsof these active compounds as a free base or pharmacologically acceptablesalt can be prepared in water suitably mixed with a surfactant such ashydroxy propylcellulose. Dispersions can also be prepared in glycerol,liquid polyethylene glycols and mixtures thereof in oils. Under ordinaryconditions of storage and use, these preparations contain a preservativeto prevent the growth of microorganisms.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. In all cases, the form must be sterile and must be fluid tothe extent that easy syringability exists. It must be stable under theconditions of manufacture and storage and must be preserved against thecontaminating action of microrganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (e.g., glycerol, propylene glycol and liquidpolyethylene glycol), suitable mixtures thereof, and vegetable oil. Theinvention will be further described in conjunction with the followingnon-limiting examples.

EXAMPLE 114-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-3-(4-fluorophenyl)-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid

To a solution of 47.3 mg (50 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioic acid λ_(max) nm in water=259) in 2.5ml of methanol is added 200 μl of pyridine and 200 μl of2,4-pentanedione at room temperature. The reaction mixture is stirredovernight (the reaction is monitored by mass spectroscopy (MS)). Afterthe reaction is complete, the volatile materials are removed underreduced pressure. N,N-dimethylformamide (DMF) (3 ml) is added to theresidue, followed by addition of 13.0 mg (95 μmol) of 4-fluorophenylisocyanate. The reaction mixture is stirred at room temperature for 50hours (the reaction was monitored by MS). The volatile materials areremoved under reduced pressure to afford a residue to which is added 5ml of 0.5% trifluoroacetic acid (TFA) in a 1:1 mixture of water andp-dioxane. The mixture is stirred at room temperature for 2 hours. Thereaction is monitored by MS and the desired product is identified byliquid chromatography/mass spectrum (LC/MS). After concentration ofvolatiles to about one ml, 24 mg (44% yield) of the desired product isobtained as a colorless solid using preparative high pressure liquidchromatography (HPLC).

HPLC conditions:

column: Prodigy ODS 4.6×150 mm

mobile phase: gradient, A=0.02% TFA/water; B=0.02%

TFA/acetonitrile

flow rate: 1.0 ml/min

detection: 215 nm, MSD

retention time: 5.1 min

Molecular formula: C₄₅H₆₅FN₁₂O₁₇;

Molecular weight: positive ion electrospray MS m/z=1065.2 (M+H)⁺ and533.2 (M+2H)²;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 1;

Ultraviolet absorption spectrum: λ_(max) nm (water)=261.

EXAMPLE 214-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-2,4-dioxo-3-pentyl-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid

The title compound is prepared by the procedure of Example 1, using 24.0mg (25 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid λ_(max) nm in water=259) in 1.0 ml of methanol, 100 μl of pyridine,100 μl of 2,4-pentanedione and 30 μmol of pentylisocyanate to give 17 mgof the desired product.

Molecular formula: C₄₄H₇₂N₁₂O₁₇;

Molecular weight: positive ion electrospray MS m/z=1041.5 (M+H)⁺ and521.4 (M+2H)²⁺.

EXAMPLE 314-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-3-hexyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid

The title compound is prepared by the procedure of Example 1, using 47.3mg (50 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) in 3.0 ml of methanol, 200 μl ofpyridine, 200 μl of 2,4-pentanedione and 50 μmol of hexylisocyanate togive 15 mg of the desired product.

HPLC conditions:

column: Prodigy ODS 4.6×150 mm

mobile phase: gradient, A=0.02% TFA/water; B=0.02%

TFA/acetonitrile

flow rate: 1.0 ml/min

detection: 215 nm, MSD

retention time: 5.3 min

Molecular formula: C₄₅H₇₄N₁₂O₁₇;

Molecular weight: positive ion electrospray MS m/z=528.4 (M+2H)²⁺.

EXAMPLE 414-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-3-(4-methoxyphenyl)-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid

The title compound is prepared by the procedure of Example 1, using 47.3mg (50 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) in 3.0 ml of methanol, 200 μl ofpyridine, 200 μl of 2,4-pentanedione and 55 μmol of4-methoxyphenylisocyanate to give 16 mg of the desired product.

HPLC conditions:

column: Prodigy ODS 4.6×150 mm

mobile phase: gradient, A=0.02% TFA/water; B=0.02%

TFA/acetonitrile

flow rate: 1.0 ml/min

detection: 215 nm, MSD

retention time: 7.31 min

Molecular formula: C₄₆H₆₈N₁₂O₁₈;

Molecular weight: positive ion electrospray MS m/z=539.5 (M+2H)²⁺.

EXAMPLE 514-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-3-dodecyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid

The title compound is prepared by the procedure of Example 1, using 56.8mg (60 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) in 3.5 ml of methanol, 240 μl ofpyridine, 240 μl of 2,4-pentanedione and 66 μmol of dodecylisocyanate togive 15 mg of the desired product.

HPLC conditions:

column: Prodigy ODS 4.6×150 mm

mobile phase: gradient, A=0.02% TFA/water; B=0.02%

TFA/acetonitrile

flow rate: 1.0 ml/min

detection: 215 nm, MSD

retention time: 10.3 min

Molecular formula: C₅₁H₈₆N₁₂O₁₇;

Molecular weight: positive ion electrospray MS m/z=1139.4 (M+H)⁺ and570.3 (M+2H)²⁺;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 2;

Ultraviolet absorption spectrum: λ_(max) nm (water)=259.

EXAMPLE 616-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-15-benzyl-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicAcid

To a solution of 56.8 mg (60 μmol)16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) in 3.5 ml of methanol is added 240 μl ofpyridine and 240 μl of 2,4-pentanedione at room temperature. Thereaction mixture is stirred for 48 hours (the reaction is monitored byMS). After the reaction is complete, the volatile materials are removedunder reduced pressure. N,N-Diemthylformamide (4 ml) is added to theresidue, followed by addition of 12.3 mg (72 μmol) of benzyl bromide.The reaction mixture is stirred at room temperature overnight (thereaction is monitored by MS). The volatile materials are removed underreduced pressure to a residue to which is added 5 ml of 0.5% TFA in a1:1 mixture of water:p-dioxane and the mixture is stirred at roomtemperature for 2 hours. The reaction is monitored by MS and the desiredproduct identified by LC/MS. After the concentration of the reactionmixture to about one ml, 7 mg of the desired product is separated bypreparative HPLC.

HPLC conditions:

column: Prodigy ODS 4.6×150 mm

mobile phase: gradient, A=0.02% TFA/water; B=0.02%

TFA/acetonitrile

flow rate: 1.0 m/min

detection: 215 nm, MSD

retention time: 5.8 min

Molecular formula: C₄₅H₆₉N₁₁O₁₇;

Molecular weight: positive ion electrospray MS m/z=1036.2 (M+H)⁺ and518.8 (M+2H)²⁺;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 3.

EXAMPLE 716-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-15-dodecyl-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicAcid

To a solution of 56.8 mg (60 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) in 3.0 ml of methanol is added 300 μl ofpyridine and 300 μl of 2,4-pentanedione at room temperature. Thereaction mixture is stirred for 2 days (the reaction is monitored byMS). After the reaction is complete, the volatile materials are removedunder reduced pressure. N,N-Dimethylformamide (3 ml) is added to theresidue, followed by addition of 14.4 mg (78 μmol) of dodecyl aldehyde.The reaction mixture is kept at 70° C. for 0.5 hour, followed byaddition of 78 μmol of sodium cyanoborohydride (NaBH₃CN), and thereaction mixture is heated at 70° C. (oil-bath) for an additional 2hours (the reaction is also monitored by MS). The volatile materials areremoved under reduced pressure to a residue to which is added 5 ml of0.5% trifluoroacetic acid in a 1:1 mixture of water:p-dioxane and thereaction mixture stirred at room temperature for 2 hours. The reactionis monitored by MS and the desired product is identified by LC/MS. Afterconcentration of the reaction mixture to about one ml, 21 mg of thedesired product is separated by preparative HPLC.

HPLC conditions:

column: Prodigy ODS 4.6×150 mm

mobile phase: gradient, A=0.02% TFA/water; B=0.02%

TFA/acetonitrile

flow rate: 1.0 ml/min

detection: 215 nm, MSD

retention time: 8.3 min

Molecular formula: C₅₀H₈₇N₁₁O₁₇;

Molecular weight: positive ion electrospray MS m/z=1115.0 (M+H)⁺ and558.2 (M+2H)²⁺;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 4;

Ultraviolet absorption spectrum: λ_(max) nm (water)=260.

EXAMPLE 816-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-15-[12-(4-morpholinyl)dodecyl]-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicAcid

To a solution of 66.2 mg (70 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) in 3.0 ml of methanol is added 300 μlpyridine, 300 μl of 2,4-pentanedione and 30 mg montmorillonite K-10 (aclay forming the principal constituent of bentonite and fuller'searth(Merck Index, 11, 6168)) at room temperature. The reaction mixtureis stirred for 2 days and monitored by MS. After the reaction iscompletion, the reaction mixture is filtered, and washed with methanol.The volatile materials are removed under reduced pressure to a residueto which is added N,N-Dimethylformamide (3 ml) followed by additon of 66mg (245 μmol) of 12-(4-morpholinyl)dodecanal (see examples 21 & 22).After stirring at room temperature for two hours, 46.8 mg of sodiumtriacetoxyborohydride and 4.2 mg of acetic acid are added. The mixtureis stirred at room temperature over the weekend (48 hr) (the reactionwas also monitored by MS) and the volatile materials are removed underreduced pressure to a residue to which is added 8 ml of 0.5%trifluoroacetic acid in a 1:1 mixture of water and p-dioxane. Thereaction mixture is stirred at room temperature for 1.5 hour. Thereaction was monitored by MS and the desired product is identified byLC/MS. The reaction mixture is concentrated to about one ml and 18.7 mg(22.3%) of the desired product is separated by preparative HPLC.

HPLC conditions:

column: Prodigy ODS 4.6×150 mm

mobile phase: gradient, A=0.02% TFA/water; B=0.02%

TFA/acetonitrile

flow rate: 1.0 ml/min

detection: 215 nm, MSD

retention time: 5.4 min

Molecular formula: C₅₄H₉₄N₁₂O₁₈;

Molecular weight: positive ion electrospray MS m/z=1199.7 (M+H)⁺ and600.7 (M+2H)²⁺; 401.0 (M+3H)³⁺;

Ultraviolet absorption spectrum: λ_(max) nm (water)=259;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 5.

EXAMPLE 916-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-15-pentyl-3,5,8,11,15-pentaazaheptadecane-1,17-dioicAcid

The title compound is prepared by the procedure of Example 8, using 56.8mg (60 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) in 3.0 ml of methanol, 300 μl ofpyridine, 300 μl of 2,4-pentanedione, 15 mg montmorillonite K-10, 18 μlof pentanal and 180 μmol of sodium triacetoxyborohydride to give 21 mgof the desired product.

HPLC conditions:

column: Prodigy ODS 4.6×150 mm

mobile phase: gradient, A=0.02% TFA/water; B=0.02%

TFA/acetonitrile

flow rate: 1.0 ml/min

detection: 215 nm, MSD

retention time: 5.7 min

Molecular formula: C₄₃H₇₃N₁₁O₁₇;

Molecular weight: positive ion electrospray MS m/z=1018.6 (M+H)⁺ and510.1 (M+2H)²⁺;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 6;

Ultraviolet absorption spectrum: λ_(max) nm (water)=259.

EXAMPLE 104-[13-((2R)-2-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}-1-carboxy-2-{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}ethyl)-26-carboxy-19-(1-hydroxy-2-methylpropyl)-22-(2-iminohexahydro-4-pyrimidinyl)-27-methyl-18,21,24-trioxo-13,17,20,23,25-pentaazaoctacos-1-yl]-4-methylmorpholin-4-ium

The title compound is prepared by the procedure of Example 8, using198.6 mg (210 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) in 3.0 ml of methanol, 300 μl ofpyridine, 300 μl of 2,4-pentanedione, 15 mg montmorillonite K-10, 18 μlof 4-methyl-4-(12-oxododecyl)morpholin-4-ium (see example 23) and 180μmol of sodium triacetoxyborohydride to give 26.7 mg of the desiredproduct.

HPLC conditions: column: Prodigy ODS 4.6×150 mm

mobile phase: gradient, A=0.02% TFA/water; B=0.02%

TFA/acetonitrile

flow rate: 1.0 ml/min

detection: 215 nm, MSD

retention time: 6.3 min

Molecular formula: C₅₅H₉₇IN₁₂O₁₈;

Molecular weight: positive ion electrospray MS m/z=809.7 (2M⁺+H)³⁺ and607.5 (M⁺+H)²⁺; 405.5 (M⁺+2H)⁺;

Ultraviolet absorption spectrum: λ_(max) nm (water)=260;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 7.

EXAMPLE 11 14-[5-((R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(3R,4S,5R)-4-hydroxy-3-methoxy-5-({[(octylamino)carbonyl]amino}methyl)tetrahydro-2-furanyl]oxy}methyl)-3-octyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicAcid

To a suspension of 47.3 mg (50 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) in 4.0 ml of N,N-dimethylformamide isadded 20.2 mg (130 μmol) of n-octyl isocyanate at room temperature. Thereaction mixture is stirred for 20 hours. The reaction is monitored byMS and checked by LC/MS. The reaction mixture is concentrated to about 1ml, and the fractions with molecular weights of 1237.9 and 1255.9 areseparated by preparative HPLC. After heating the mixture in 5 ml ofwater at 70° C. for 45 minutes, 27 mg (43.6%) of the desired product(molecular weight=1237.9) is obtained.

HPLC conditions:

column: Prodigy ODS 4.6×150 mm

mobile phase: gradient, A=0.02% TFA/water; B=0.02%

TFA/acetonitrile

flow rate: 1.0 ml/min

detection: 215 nm, MSD

Molecular formula: C₅₆H₉₅N₁₃O₁₈;

Molecular weight: positive ion electrospray MS m/z=1238.85 (M+H)⁺ and619.86 (M+2H)²⁺;

Ultraviolet absorption spectrum: λ_(max) nm (water)=262;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 8.

EXAMPLE 12 14-[5-((R)-{(2S,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(5R)-4-hydroxy-3-methoxy-5-({[(4-fluoroanilino)carbonyl]amino}methyl)tetrahydro-2-furanyl]oxy}methyl)-3-(4-fluorophenyl)-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicAcid

The title compound is prepared by the procedure of Example 11, using47.3 mg (50 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) and 27.4 mg (200 μmol) 4-methoxyphenylisocyanate in 4.0 ml of N,N-dimethyl formamide to give 17.3 mg of thedesired product.

Molecular formula: C₅₂H₆₉F₂N₁₃O₁₈;

Molecular weight: positive ion electrospray MS m/z=1202.7 (M+H)⁺ and601.6 (M+2H)²⁺;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 9.

EXAMPLE 13 14-[5-((R)-{(2S,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(5R)-4-hydroxy-3-methoxy-5-({[(4-methoxyanilino)carbonyl]amino}methyl)tetrahydro-2-furanyl]oxy}methyl)-3-(4-methoxyphenyl)-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicAcid

The title compound is prepared by the procedure of Example 11, using47.3 Mg (50 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) and 14.9 mg (100 μmol) 4-methoxyphenylisocyanate in 4.0 ml of N,N-dimethyl formamide to give 28.6 mg of thedesired product.

Molecular formula: C₅₄H₇₅IN₁₃O₂₀;

Molecular weight: positive ion electrospray MS m/z=1226.7 (M+H)⁺ and613.7 (M+2H)²⁺.

EXAMPLE 14 14-[5-((R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(3R,4S,5R)-5-({[(hexylamino)carbonyl]amino}methyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}methyl)-3-hexyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicAcid

The title compound is prepared by the procedure of Example 11, using47.3 mg (50 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) and 16.5 mg (130 μmol) N-hexyl isocyanatein 4.0 ml of N,N-dimethyl formamide to give 16.4 mg of the desiredproduct.

Molecular formula: C₅₂H₈₇N₁₃O₁₈;

Molecular weight: positive ion electrospray MS m/z=1182.6 (M+H)⁺ and591.7 (M+2H)²⁺;

Ultraviolet absorption spectrum: λ_(max) nm (water)=262;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 10.

EXAMPLE 15 14-[5-((R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(3R,4S,5R)-5-({[(dodecylamino)carbonyl]amino}methyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}methyl)-3-dodecyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicAcid

The title compound is prepared by the procedure of Example 11, using56.8 mg (60 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) and 14.9 mg (100 μmol) n-dodecylisocyanate in 4.0 ml of N,N-dimethyl formamide to give 30.0 mg of thedesired product.

Molecular formula: C₆₄H₁₁₁N₁₃O₁₈;

Molecular weight: positive ion electrospray MS m/z=1350.6 (M+H)⁺ and675.8 (M+2H)²⁺.

EXAMPLE 16 16-((R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(3R,4S,5R)-5-({[(hexadecylamino)carbonyl]amino}methyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicAcid

The title compound is prepared by the procedure of Example 11, using50.0 mg (52 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) and 14.0 mg (52 μmol) hexadecylisocyanate in 2.0 ml of N,N-dimethyl formamide to give 3.0 mg of thedesired product.

Molecular formula: C₅₅H₉₆N₁₂O₁₈;

Molecular weight: positive ion electrospray MS m/z=1213.5 (M+H)⁺ and607.4 (M+2H)²⁺;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 11.

EXAMPLE 17 16-[(R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}({(3R,4S,5R)-4-hydroxy-3-methoxy-5-[(pentylamino)methyl]tetrahydro-2-furanyl}oxy)methyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-15-pentyl-3,5,8,11,15-pentaazaheptadecane-1,17-dioicAcid

To a solution of 50 mg (53 μmol)16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) in 0.4 ml of methanol is added 9.5 mg(2.0 eq., 110 μmol) of valeraldehyde and 3.3 mg (1.0 eq., 53 μmol) ofsodium cyanoborohydride. After stirring for 5 minutes, 0.04 ml of 1 Nmethanolic hydrochloric acid is added. The reaction mixture is thenstirred for 0.5 hour (the reaction is monitored by MS). The solvent isremoved under reduced pressure to give a yellow gum. The desired productis identified by LC/MS and separated by preparative HPLC to give 4.9 mg(9.0% yield) of the desired product as a white solid.

Molecular formula: C₄₈H₈₃N₁₁ O₁₇;

Molecular weight: positive ion electrospray MS m/z=1086.7 (M+H)⁺ and543.8 (M+2H)²⁺.

EXAMPLE 18 16-[(R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}({(3R,4S,5R)-4-hydroxy-3-methoxy-5-[(pentylamino)methyl]tetrahydro-2-furanyl}oxy)methyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicAcid

The title compound is prepared by the procedure of Example 17, using50.0 mg (53 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) in 0.4 ml of methanol, 4.7 mg (1.0 eq.,55 μmol) of valeraldehyde and 3.3 mg (1.0 eq., 53 μmol) of sodiumcyanoborohydride to give 7.0 mg of the desired product.

Molecular formula: C₄₃H₇₃N₁₁O₁₇;

Molecular weight: positive ion electrospray MS m/z=1016 (M+H)⁺ and 508.8(M+2H)²⁺;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 12.

EXAMPLES 19a AND 19b16-((R)-[((3R,4S,5R)-5-{[([1,1′-Biphenyl]-4-ylmethyl)amino]methyl}-4-hydroxy-3-methoxytetrahydro-2-furanyl)oxy]{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicAcid (Example 19a) and15-([1,1′-Biphenyl]-4-ylmethyl)-16-((R)-[((3R,4S,5R)-5-{[([1,1′-biphenyl]-4-ylmethyl)amino]methyl}-4-hydroxy-3-methoxytetrahydro-2-furanyl)oxy]{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicAcid (Example 19b)

To a solution of 20 mg (21 μmol)16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) in 0.4 ml of methanol is added 77 mg (2.0eq., 42 μmol) of 4-biphenylcarboxaldehyde and 1.3 mg (1.0 eq., 21 μmol)of sodium cyanoborohydride. After stirring for 5 minutes, 0.04 ml ofmethanolic hydrochloric acid is added. The reaction mixture is thenstirred for 0.5 hour and monitored by MS. The solvent is removed underreduced pressure to give a yellow gum. The desired products areseparated by preparative HPLC to give 3.0 mg (12% yield) of Example 19aas a white solid and 4.0 mg (15% yield, L17742-171-2) of Example 19balso as a white solid.

Example 19a

Molecular formula: C₅₁H₇₃N₁₁O₁₇;

Molecular weight: positive ion electrospray MS m/z=1112.5 (M+H)⁺ and556.7 (M+2H)²⁺;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 13.

Example 19b

Molecular formula: C₆₄H₈₃N₁₁O₁₇;

Molecular weight: positive ion electrospray MS m/z=1278.7 (M+H)⁺ and639.8 (M+2H)²⁺;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 14.

EXAMPLE 20a and 20b16-((R)-({(3R,4S,5R)-5-[(Benzylamino)methyl]-4-hydroxy-3-methoxytetrahydro-2-furanyl}oxy){(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (Example 20a) and15-Benzyl-16-((R)-({(3R,4S,5R)-5-[(benzylamino)methyl]-4-hydroxy-3-methoxytetrahydro-2-furanyl}oxy){(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicAcid (Example 20b)

To a solution of 50 mg (53 μmol)16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid (λ_(max) nm in water=259) in 0.4 ml of methanol is added 11 mg (2.0eq., 106 μmol) of benzaldehyde and 3.3 mg (1.0 eq., 53 μmol) of NaBH₃CN.After stirring for 5 minutes, 0.04 ml of methanolic hydrochloric acid isadded. The reaction mixture is then stirred for 0.5 hour. The reactionis monitored by MS. The solvent is removed under reduced pressure togive a yellow gum. The desired products are separated by preparativeHPLC to give 15 mg (26% yield) of Example 20a as a white solid and 2.5mg (4% yield) of Example 20b as a white solid.

Example 20a

Molecular formula: C₄₅H₆₉N₁₁O₁₇;

Molecular weight: positive ion electrospray MS m/z=1036.1 (M+H)⁺ and518.9 (M+2H)²⁺;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 15.

Example 20b

Molecular formula: C₅₂H₇₅N₁₁O₁₇;

Molecular weight: positive ion electrospray MS m/z=1126.5 (M+H)⁺ and563.8 (M+2H)²⁺;

Proton magnetic resonance spectrum (300 Mhz D₂O): FIG. 16.

EXAMPLE 21 12-(4-Morpholinyl)-1-dodecanol

To a solution of 5.30 g (20 mmol) of 12-bromo-1-dodecanol in 100 ml ofacetonitrile is added dropwise 6 ml (68.6 mmol) of morpholine at 0° C.The solution is then stirred at room temperature overnight and filtered.After removal of the acetonitrile, the residue is dissolved in 250 ml ofdichloromethane. The dichloromethane solution is washed with water,dried over magnesium sulfate and filtered. The dichloromethane isremoved under reduced pressure and the residue is purified by silica gelcolumn chromatography to give 4.2 g (79.2%) of12-(4-morpholinyl)-1-dodecanol.

Molecular formula: C₁₆H₃₃NO₂;

Proton magnetic resonance spectrum (300 Mhz CH₃Cl): 3.72 (t, 4H, J=4.6Hz); 3.66 (t, 2H, J=6.5 Hz); 2.43 (t, 4H, J=4.5 Hz); 2.34 (t, 2H, J=6.9Hz); 1.50 (m, 21H);

Molecular weight: positive ion electrospray MS m/z=272.4 (M+H)⁺;

Elemental analysis: theory, C 70.80; H 12 25; N 5.16; found, C 71.12; H12.32; N 5.11.

EXAMPLE 22 12-(4-Morpholinyl)-1-dodecanal

A mixture of 542.9 mg (2.0 mmol) of 12-(4-morpholinyl)-1-dodecanol and3.0 g of (5.10 mmol) of bis(trimethylsilyl)chromate (M. Heravi, Monatsh.Chem. 1998, 129(12), 1305-1308) in 10 ml of dichloromethane is stirredat room temperature overnight and filtered. After removal of thedichloromethane, the residue is purified by silica gel columnchromatography to give 210 mg (39%) of 12-(4-morpholinyl)-1-dodecanal.

Molecular formula: C₁₆H₃₁NO₂;

Proton magnetic resonance spectrum (300 Mhz CH₃Cl): 9.76 (t, 1H, J=1.9Hz); 3.72 (t, 4H, J=4.7 Hz); 2.43 (t, 4H, J=4.5 Hz); 2.29 (t, 2H, J=5.3Hz); 1.50 (m, 20H);

Molecular weight: positive ion electrospray MS m/z=270.3 (M+H)⁺.

EXAMPLE 23 4-Methyl-4-(12-oxododecyl)morpholin-4-ium

A mixture of 1.0 g (3.7 mmol) of 12-(4-morpholinyl)-1-dodecanal and 4.26g of (1.86 ml; 30 mmol) of iodomethane in 20 ml of diethyl ether isstirred at room temperature overnight.4-Methyl-4-(12-oxododecyl)morpholin-4-ium which precipitated from thesolution is collected by filtration, washed with diethyl ether and driedto give 259 mg of the desired product.

Molecular formula: C₁₇H₃₄NO₂;

Molecular weight: positive ion electrospray MS m/z=284.4 M⁺.

EXAMPLE 2416-((R)-({(3R,4S,5R)-5-[(Acetamido)methyl]-4-hydroxy-3-methoxytetrahydro-2-furanyl}oxy){(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicAcid

A suspension of 39.0 mg (41.2 μmol) of16-({[5-(aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid in 2.0 ml of acetic anhydride is placed in an ultrasonic bath(Branson 1200) at room temperature for 35 minutes. Water (2 ml) andn-butanol (2 ml) are added to the reaction mixture and the volatilematerials removed under reduced pressure to a residue which is suspendedin 2 ml of acetone and the suspension filtered through glass wool. Thedesired product (the acetone insoluble product) is recovered as a whitesolid by dissolving in 2 ml water followed by lyophylization.

Molecular formula: C₄₀H₆₅N₁₁O₁₈;

Molecular weight: positive ion electrospray MS m/z=988.4 (M+H)⁺ and494.7 (M+2H)²⁺;

Proton magnetic resonance spectrum (300 MHz D₂O): FIG. 17.

We claim:
 1. A compound of the formula:

wherein: R₁ is H, aryl, alkyl (C₁-C₂₀), —CH₂-aryl, —C(O)alkyl(C₁-C₂₀),—C(O)NHalkyl(C₁-C₂₀), or —C(O)NHaryl; R₂ is H, alkyl (C₁-C₂₀), —CH₂aryl,alkyl (C₁-C₂₀) or —C(O)alkyl(C₁-C₂₀); R₃ is —OH; R₂ and R₃ mayoptionally be taken together to form a moiety

R₄ is alkyl (C₁-C₂₀), or aryl; provided R₁ and R₂ are not H when R₃ is—OH or a pharmaceutically acceptable salt thereof.
 2. A compoundaccording to claim 1 wherein: R₂ is H, alkyl (C₁-C₁₂), or —CH₂aryl.
 3. Acompound according to claim 1 wherein: R₁ is H, —C(O)alkyl(C₁-C₁₆), or—C(O)aryl when R₂ and R₃ are taken together to form a moiety


4. A compound according to claim 1 wherein: R₄ is alkyl(C₁-C₁₆), oraryl.
 5. The compound according to claim 1 which is4-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-3-(4-fluorophenyl)-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid.
 6. The compound according to claim 1 which is14-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-2,4-dioxo-3-pentyl-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid.
 7. The compound according to claim 1 which is14-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-3-hexyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid.
 8. The compound according to claim 1 which is14-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-3-(4-methoxyphenyl)-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid.
 9. The compound according to claim 1 which is14-[5-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-3-dodecyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid.
 10. The compound according to claim 1 which is16-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-15-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid.
 11. The compound according to claim 1 which is16-((R)-{[(3R,4S,5R)-5-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-15-dodecyl-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid.
 12. The compound according to claim 1 which is16-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-15-[12-(4-morpholinyl)dodecyl]-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid.
 13. The compound according to claim 1 which is16-((R)-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-15-pentyl-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid.
 14. The compound according to claim 1 which is4-[13-((2R)-2-{[(3R,4S,5R)-5-(Aminomethyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}-1-carboxy-2-{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}ethyl)-26-carboxy-19-(1-hydroxy-2-methylpropyl)-22-(2-iminohexahydro-4-pyrimidinyl)-27-methyl-18,21,24-trioxo-13,17,20,23,25-pentaazaoctacos-1-yl]-4-methylmorpholin-4-ium.15. The compound according to claim 1 which is14-[5-((R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(3R,4S,5R)-4-hydroxy-3-methoxy-5-({[(octylamino)carbonyl]amino}methyl)tetrahydro-2-furanyl]oxy}methyl)-3-octyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid.
 16. The compound according to claim 1 which is14-[5-((R)-{(2S,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(5R)-4-hydroxy-3-methoxy-5-({[(4-fluoroanilino)carbonyl]amino}methyl)tetrahydro-2-furanyl]oxy}methyl)-3-(4-fluorophenyl)-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid.
 17. The compound according to claim 1 which is14-[5-((R)-{(2S,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(5R)-4-hydroxy-3-methoxy-5-({[(4-methoxyanilino)carbonyl]amino}methyl)tetrahydro-2-furanyl]oxy}methyl)-3-(4-methoxyphenyl)-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid.
 18. The compound according to claim 1 which is14-[5-((R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(3R,4S,5R)-5-({[(hexylamino)carbonyl]amino}methyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}methyl)-3-hexyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid.
 19. The compound according to claim 1 which is14-[5-((R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(3R,4S,5R)-5-({[(dodecylamino)carbonyl]amino}methyl)-4-hydroxy-3-methoxytetrahydro-2-furanyl]oxy}methyl)-3-dodecyl-2,4-dioxo-1-imidazolidinyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11-tetraazatetradecan-1-oicacid.
 20. The compound according to claim 1 which is16-((R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}{[(3R,4S,5R)-furanyl]oxy}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid.
 21. The compound according to claim 1 which is16-[(R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}({(3R,4S,5R)-4-hydroxy-3-methoxy-5-[(pentylamino)methyl]tetrahydro-2-furanyl}oxy)methyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-15-pentyl-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid.
 22. The compound according to claim 1 which is16-[(R)-{(2S,3S,4R,5R)-5-[2,4-Dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}({(3R,4S,5R)-4-hydroxy-3-methoxy-5-[(pentylamino)methyl]tetrahydro-2-furanyl}oxy)methyl]-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid.
 23. The compound according to claim 1 which is16-((R)-[((3R,4S,5R)-5-{[([1,1′-Biphenyl]-4-ylmethyl)amino]methyl}-4-hydroxy-3-methoxytetrahydro-2-furanyl)oxy]{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid.
 24. The compound according to claim 1 which is15-([1,1′-Biphenyl]-4-ylmethyl)-16-((R)-[((3R,4S,5R)-5-{[([1,1′-biphenyl]-4-ylmethyl)amino]methyl}-4-hydroxy-3-methoxytetrahydro-2-furanyl)oxy]{(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid.
 25. The compound according to claim 1 which is16-((R)-({(3R,4S,5R)-5-[(Benzylamino)methyl]-4-hydroxy-3-methoxytetrahydro-2-furanyl}oxy){(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid.
 26. The compound according to claim 1 which is15-Benzyl-16-((R)-({(3R,4S,5R)-5-[(benzylamino)methyl]-4-hydroxy-3-methoxytetrahydro-2-furanyl}oxy){(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid.
 27. The compound according to claim 1 which is16-((R)-({(3R,4S,5R)-5-[(Acetamido)methyl]-4-hydroxy-3-methoxytetrahydro-2-furanyl}oxy){(2S,3S,4R,5R)-5-[2,4-dioxo-3,4-dihydro-1(2H)-pyrimidinyl]-3,4-dihydroxytetrahydro-2-furanyl}methyl)-9-(1-hydroxy-2-methylpropyl)-6-(2-iminohexahydro-4-pyrimidinyl)-2-isopropyl-4,7,10-trioxo-3,5,8,11,15-pentaazaheptadecane-1,17-dioicacid.
 28. A method for treating bacterial infections in warm bloodedanimals which comprises providing to said animals an antibacteriallyeffective amount of a compound according to claim
 1. 29. Apharmaceutical composition which comprises a compound according to claim1 in association with a pharmaceutically acceptable carrier.
 30. Aprocess for the preparation of an AA-896 antibiotic of the formula

where R₄ is alkyl (C₁-C₂₀), or aryl; which comprises: a. protecting theprimary amine of

 with 2,4-pentanedione in pyridine:methanol to form a protected amine;b. reacting the secondary amine with an isocyanate, R₄NCO; and c.removal of the primary amine protecting group with aqueoustrifluoroacetic acid to form an AA-896 antibiotic.
 31. A process for thepreparation of an AA-896 antibiotic of the formula

where R₂ is alkyl (C₁-C₁₂), or —CH₂aryl; which comprises: a. protectingthe primary amine of

 with 2,4-pentanedione in pyridine:methanol to form a protected amine;b. reacting the secondary amine with an aldehyde, R₄CH═O in the presenceof a reducing agent such as sodium cyanoborohydride or sodiumtriacetoxyborohydride; and c. removal of the primary amine protectinggroup with aqueous trifluoroacetic acid to form an AA-896 antibiotic.